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Seed Fatty Acid Compositions and Chemotaxonomy of Wild Crambe (Brassicaceae) Taxa in Turkey

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Seed Fatty Acid Compositions and Chemotaxonomy of Wild Crambe (Brassicaceae) Taxa in Turkey Turkish Journal of Agriculture and Forestry Turk J Agric For (2020) 44: 662-670 http://journals.tubitak.gov.tr/agriculture/ © TÜBİTAK Research Article doi:10.3906/tar-1912-76 Seed fatty acid compositions and chemotaxonomy of wild Crambe (Brassicaceae) taxa in Turkey İlhan SUBAŞI* Central Research Institute for Field Crops, Ankara, Turkey Received: 31.12.2019 Accepted/Published Online: 01.09.2020 Final Version: 08.12.2020 Abstract: Wild Crambe species have greater potential than Crambe hispanica in industry, medicine, as a vegetable, etc. A total of 53 germplasm accessions, belonging to 7 taxa, were collected from the natural flora of Turkey. The accessions consisted ofC. orientalis var. orientalis (18 accessions), C. orientalis var. dasycarpa (1 accession), C. orientalis var. sulphurea (2 accessions), C.tataria var. aspera (3 accessions), C. tataria var. tataria (26 accessions), C. grandiflora (1 accession), C. orientalis var. sulphurea, and C. maritima (2 accessions). In this study, the seed fatty acid compositions and oil contents were determined, and the data were used for taxonomic cluster, correlation, and principal component analyses. Important correlations were determined among the fatty acids; however, the oil contents were not correlated. Altitude was positively correlated with linolenic acid, while negatively correlated with oleic and linoleic acid. For the principal component and correlation analyses, 7 major fatty acids (>1%) were used, including palmitic (C16:0), oleic (C18:1), linoleic (C18:2), cis-11 eicosenoic (20:1), linolenic (C18:3), erucic (C22:1), and nervonic acid (C24:1). A total of 17 fatty acids were used for the cluster analyses. Two major clusters were formed, where the first consisted of C. orientalis, C. tataria, and C. grandiflora taxa, while the second consisted of only C. maritima taxa. The dendrogram based on the fatty acid values clearly discriminated the species groups; however, C. tataria was not located close to C. maritima, contrary to previous molecular cluster studies. Key words: Chemotaxonomy, fatty acid composition, seed oil content, wild Crambe 1. Introduction The oil content of cultivated C. abyssinica species Indigenous plants form the bedrock of diversity in has been reported to have a wide range of variation, industrial systems. Deficiently used industrial crop from 24% to 46% (Warwick and Black, 1997; Gastaldi et genetic resources have great potential as an alternative to al., 1998; Comlekcioglu et al., 2008; Rudloff and Wang, worldwide known species or varieties that are commonly 2011; Uyaroğlu, 2018). The total fatty acid composition produced and consumed. With climate uncertainty, there comprised 53.9%–63.1% erucic acid (Vollmann and is an urgent need to diversify plant resources to a wider Ruckenbauer, 1993). Little information on the oil content range of crops for greater system resistance (Zia-Ul-Haq et of other wild Crambe species has been reported. The al., 2013; Colak et al., 2019). whole-seed oil content was reported as 11% and 15% in The genus Crambe comprises annual, biennial, and C. orientalis and C. tataria, respectively (Comlekcioglu perennial species (Comlekcioglu et al., 2008). It has et al., 2008). However, although the reported oil and potential in terms of use in the oleochemical industry; erucic acid rates of wild Crambe species have been low, however, currently, C. abyssinica Hochst. is the only they are a significant source of genetic variation for fatty species of Crambe that is used in industry (Warwick and acid composition. Wild Crambe seed oil mainly contains Black, 1997). Due to its high amount of erucic acid, which palmitic, oleic, cis-11 eicosenoic, erucic, linoleic, and provides high boiling and evaporating points, it is widely linolenic acid. used in industrial applications, such as the production of Understanding the genetic potential and characteristics plastic film, nylon, coatings, and lubricants (Warwick and of wild relatives of the different plant species is important Black, 1997; Piovan et al., 2011). for long-term breeding programs (Ersoy et al., 2018; There are 6 species and 4 intraspecific taxa in the Gecer et al., 2020). In addition, many wild species have natural flora of Turkey, while there are 35 taxa worldwide the potential to become new valuable crop plants. As (Prina, 2009; Mutlu, 2012; Bassegio et al., 2016; Tarıkahya- an industrial oil source, wild relatives of Crambe may Hacıoğlu, 2016). possess many important agronomical properties, such * Correspondence: [email protected] 662 This work is licensed under a Creative Commons Attribution 4.0 International License. SUBAŞI / Turk J Agric For as cytoplasmic and nuclear male infertility, disease and were set at 250 °C. The column was programmed with the pest resistance, C3-C4 photosynthetic activity, and cold, following temperature regime: hold at 140 °C for 5 min, drought, and salinity resistance; therefore, they could be ramp to 240 °C at 4 °C/min, and hold at 240 °C for 20 used in breeding programs (Warwick et al., 2009). min. Fatty acid methyl esters (FAMEs) were identified In this study, it was aimed to determine the seed oil by comparison of their retention times with those of the properties of some wild Crambe accessions and investigate reference standards (Restek FAME Mix 37). The fatty acid whether the seed oil compositions could be used as a content was calculated based on the peak area ratio and taxonomical character for different Crambe species and expressed as grams of fatty acid/100 g of oil. taxa. 2.3. Statistical analysis All statistical analyses were performed using JMP Pro 11 2. Materials and methods (SAS Institute Inc., Cary, NC, USA, 2013). Correlation 2.1. Material among the studied traits was calculated using the Pearson Crambe seeds were obtained from multiple flora collection correlation procedure implemented in the mentioned expeditions in 2014 in Turkey. Among these, 53 germplasm statistical program. Principal component analysis (PCA) accessions were chosen to represent 7 Crambe taxa was performed using the fatty acid (palmitic, oleic, linoleic, collected from different regions of Turkey, comprising18 of linolenic, cis-11 eicosenoic, and erucic acids) values of C. orientalis var. orientalis, 1 of C. orientalis var. dasycarpa, each of the 7 accessions. For chemotaxonomic evaluation 2 of C.orientalis var. sulphurea, 3 of C. tataria var. aspera, of the 7 Crambe taxa, the mean values of the 17 fatty acids 1 of C. grandiflora, 21 of C. tataria var. tataria, and 2 of C. were used to perform the cluster analyses. maritima. Each taxon is listed in Table 1. The distribution of the species in Turkey is given in Figure 1.C. orientalis 3. Results was collected at an altitude of 800–1897 m, C. tataria at Within the wild relative species of Crambe, 17 fatty acids 744–1347 m, and C. maritima at 0–4 m. were determined, of which the 7 main fatty acids (>1%) were palmitic (C16:0), oleic (C18:1), linoleic (C18:2), cis- 2.2. Extraction and gas chromatography 11 eicosenoic (20:1), linolenic (C18:3), erucic (C22:1), and The oil contents and compositions were determined in nervonic (C24:1) (Table 2). Acids identified with <1% of the the laboratories of the Central Research Institute for total fatty acid content were myristic (C14:0), palmitoleic Field Crops of the Ministry of Agriculture and Forestry, (C16:1), stearic (C18:0), arashidic (C20:0), heneicosanoic in Ankara, Turkey. Determination of the oil contents was (C21:0), cis-11,14-eicosadienoic (C20:2), behenic (C22:0), performed following the method of Matthäus and Brühl cis-11,14-eicosadienoic (C20:2), cis-13,16-docosadienoic (2001), with some modifications. First, 5 grams of whole (C22:2), and lignoceric (C24:0) (Table 3). seeds were milled in a Foss KN 195 Knifetec laboratory The maximum and minimum oil contents for palmitic mill (Hillerød, Denmark). Afterwards, the extraction was (C16:0), oleic (C18:1), linoleic (C18:2), cis-11 eicosenoic performed for 3 h using a Soxhlet extractor (Soxtherm 2000 (C20:1), linolenic (C18:3), erucic (C22:1), and nervonic automatic, C. Gerhardt GmbH & Co. KG, Königswinter, (C24:1) acid were determined as 4.70%–20.50%, 0.17%– Germany) with hexane. After extraction, the solvent was 5.78%, 15.02%–32.49%, 3.94%–17.45%, 10.76%–21.79%, evaporated at 103 °C for 2.5 h, cooled for 30 min in a 2.18%–9.57%, 22.59%–49.68%, and 0.79%–2.86% in desiccator, and weighed. The seed oil content was reported all of the Crambe accessions, respectively (Table 2). C. as the mass percent [crude oil weight (g)/ground seeds orientalis accessions were identified as possessing higher weight (g) × 100]. mean values of erucic acid than the other Crambe species For determination of the fatty acid compositions, the accessions. method given by Sampaio et al. (2012) was followed, with According to the biplot diagram, 71% of the total some modifications. First, 0.1 g of oil was dissolved in 10 mL variation was explained (Figure 2). The first 4 eigenvalues of n-hexane, to which 0.5 mL of 2 N potassium hydroxide/ were 3.57, 1.47, 0.72, and 0.59. For taxonomic evaluation, methanol solution was added, and then incubated for 30 the mean values of all of the determined seed fatty acids min at room temperature. The fatty acids were esterified were used. As in the cluster analyses, all accessions as methyl esters and placed into a Shimadzu AOC-20i belonging to the same species revealed close associations. automatic injector (Shimadzu Corp., Kyoto, Japan) (split There were 4 species groups, comprisingC. orientalis, C. ratio 1:100). The methyl ester phase was analyzed using a grandiflora, C. tataria, and C. maritima. C. grandiflora was Shimadzu GC-2010 system equipped with a Teknokroma located in the C. orientalis accession group, while the C.
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